Improved detection of variants in recombinant human interferon alpha-2a products by reverse-phase high-performance liquid chromatography on a core-shell stationary phase

被引：8
作者：
Li, Yonghong [1 ,2 ]
Rao, Chunming [1 ,2 ]
Tao, Lei [1 ,2 ]
Wang, Junzhi [1 ,2 ]
Lorbetskie, Barry [3 ]
Girard, Michel [3 ]
机构：
[1] Natl Inst Food & Drug Control, Key Lab Minist Hlth Res Qual & Standardizat Biote, Beijing 100050, Peoples R China
[2] Natl Inst Food & Drug Control, WHO Collaborating Ctr Standardizat & Evaluat Biol, Beijing 100050, Peoples R China
[3] Hlth Canada, Biol & Genet Therapies Directorate, Ottawa, ON K1A 0K9, Canada
来源：
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS | 2014年 / 88卷
关键词：
Interferon alpha-2a; Variants; HPLC; Quality control; Core-shell; N-TERMINUS; STABILITY; PROTEINS; THERAPY; RESIDUE; YIELD;
D O I：
10.1016/j.jpba.2013.08.011
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
The detection of variants is one of the important aspects in quality control of recombinant DNA drugs. In this study, a gradient reverse-phase high-performance liquid chromatography (RP-HPLC) method with fluorescence detection is described for the separation of interferon alpha-2a (rhIFN alpha-2a) from several product related variants. The methodology employed a core-shell C18 column with a linear gradient elution of 0.2% (v/v) trifluoroacetic acid (TFA)-acetonitrile (ACN) at 1.0 mL/min, and the temperature of the column was maintained at 60 C. The method was validated in terms of linearity, sensitivity, intra- and inter-day variations. Compared to the European Pharmacopeia RP-HPLC method of rhIFN a-2a analysis, this new method can separate N-methionylated variant in both drug substance and finished product, and analyze the variants in untreated, oxidized sample and slightly degraded samples more efficiently. In conclusion the method has an improved capability to detect variants in rhIFN alpha-2a products. Crown Copyright (C) 2013 Published by Elsevier B.V. All rights reserved.
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收藏
页码：123 / 129
页数：7
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